Fleischmajer R, Contard P, Schwartz E, MacDonald E D, Jacobs L, Sakai L Y
Department of Dermatology, Mount Sinai School of Medicine, New York, New York 10029.
J Invest Dermatol. 1991 Oct;97(4):638-43. doi: 10.1111/1523-1747.ep12483132.
The purpose of this study is to present a three-dimensional dermal fibroblast model. Skin fibroblasts cultured in this system deposit large amounts of collagen and microfibrils. Fibroblasts were seeded onto a nylon filtration mesh and incubated in the presence or absence of ascorbic acid. Collagen fibril formation was found in the presence of ascorbic acid whereas microfibril formation was seen independent of ascorbic acid supplementation. Immunoelectron microscopy revealed that microfibrils were labeled with fibrillin at 67 nm periodicity. Isolated microfibrils studied by rotary shadowing had a beaded appearance consisting of beads linked to each other by a filamentous structure. The spaces between the beads ranged from 10.00-33.33 nm, suggesting that these microfibrils may have an extension-contraction mechanism. Furthermore, the size and spacing of the beads were similar to that seen in microfibrils from tissues (measured after rotary shadowing). Fibroblasts cultured in a three-dimensional mesh represent an effective in vitro model with which to study microfibril formation.
本研究的目的是呈现一种三维真皮成纤维细胞模型。在此系统中培养的皮肤成纤维细胞会沉积大量的胶原蛋白和微原纤维。将成纤维细胞接种到尼龙滤网上,并在有或没有抗坏血酸的情况下进行培养。在有抗坏血酸存在的情况下发现有胶原纤维形成,而微原纤维的形成则与抗坏血酸的添加无关。免疫电子显微镜显示,微原纤维以67纳米的周期被原纤蛋白标记。通过旋转投影研究的分离微原纤维具有串珠状外观,由通过丝状结构相互连接的珠子组成。珠子之间的间距在10.00 - 33.33纳米之间,这表明这些微原纤维可能具有伸展 - 收缩机制。此外,珠子的大小和间距与组织中的微原纤维(旋转投影后测量)相似。在三维网中培养的成纤维细胞代表了一种用于研究微原纤维形成的有效体外模型。